Callus induction through Anther and Ovary of Kenaf (Hibiscus cannabinus L.)

In this research new protocol has been developed that would induce callus from
anther and ovary of Kenaf. The haploid technique is a fast and efficient tool for
developing new varieties in a comparatively short time. The present study was
carried out to investigate the effect of cold treatment, plant growth regulators
(PGR) and culture condition on callus induction from Kenaf anther and ovary
culture. Kenaf HF992 cultivar was chosen as an explants material, several trails
were carried out to investigate the Androgenesis and gynogenesis ability before
we succeed to get high percentage of callus. Flower buds at the appropriate stage
of anther and ovary development were sterilized and the anthers and ovary were
carefully excised from the flowers and underwent to various pretreatments and
inoculated into media contained different combinations of PGR like NAA(anaphthaleneacetic
acid), BAP(N6-benzyladenine), 2iP(N6-(2-Isopentenyl)
adenine) and TDZ (Thidiazuron)and kept in the dark place for different periods
before transferred to light conditions. The best callus induction frequency from
anther was 90.00 % in the optimized MS medium fortified with 3.0 mg/l 2iP +
3.0 mg/l NAA at the stage the microspores were at the uninucleate stage and the
anther was about 7±1 mm long collected. And the best callus induction
frequency from ovary was 91.25% in the optimized semi solid MS medium
fortified with 3.0 mg/l 2iP + 3.0 mg/l NAA, and the flower buds was about
24±1 mm length which was collected 3-5 weeks after flower initiation. The
effect of culture condition was highly significant, the root induction was highest
(83.75% & 87.50%) of anther and ovary respectively when it kept in dark place
for 28 days.